scholarly journals Cardiac myocyte gene expression profiling during H2O2-induced apoptosis

2007 ◽  
Vol 29 (2) ◽  
pp. 118-127 ◽  
Author(s):  
Angela Clerk ◽  
Timothy J. Kemp ◽  
Georgia Zoumpoulidou ◽  
Peter H. Sugden
Keyword(s):  
Gene Expression ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Apoptotic Response ◽  
Dynamic Changes ◽  
Affymetrix Microarrays ◽  
Low Concentrations ◽  
Alternatively Spliced ◽  
Induced Apoptosis ◽  
Cardiac Myocyte Apoptosis

High levels of oxidative stress promote cardiac myocyte death, though lower levels are potentially cytoprotective/anabolic. We examined the changes in gene expression in rat neonatal cardiac myocytes exposed to apoptotic (0.2 mM) or nontoxic (0.04 mM) concentrations of H2O2 (2, 4, or 24 h) using Affymetrix microarrays. Using U34B arrays, we identified a ubiquitously expressed, novel H2O2-responsive gene [putative peroxide-inducible transcript 1 (Perit1)], which generates two alternatively spliced transcripts. Using 230 2.0 arrays, H2O2 (0.04 mM) promoted significant changes in expression of only 32 genes, all of which were seen with 0.2 mM H2O2. We failed to detect any increase in the rate of protein synthesis in cardiac myocytes exposed to <0.1 mM H2O2, further suggesting that global, low concentrations of H2O2 are not anabolic in this system. H2O2 (0.2 mM) promoted significant ( P < 0.05, >1.75-fold) changes in expression of 649 mRNAs and 187 RNAs corresponding to no established gene. Of the mRNAs, 114 encoded transcriptional regulators including Krüppel-like factors (Klfs). Quantitative PCR independently verified the changes in Klf expression. Thus, H2O2-induced cardiac myocyte apoptosis is associated with dynamic changes in gene expression. The expression of these genes and their protein products potentially influences the progression of the apoptotic response.

Gene expression changes associated with fibronectin-induced cardiac myocyte hypertrophy

2004 ◽  
Vol 18 (3) ◽  
pp. 273-283 ◽  
Author(s):  
Hua Chen ◽  
Xueyin N. Huang ◽  
Alexandre F. R. Stewart ◽  
Jorge L. Sepulveda
Keyword(s):  
Gene Expression ◽  
Cardiac Myocytes ◽  
Matrix Protein ◽  
Cardiac Myocyte ◽  
Tissue Growth ◽  
In Vivo Studies ◽  
Extracellular Matrix Protein ◽  
Myocyte Hypertrophy ◽  
Cardiac Myocyte Hypertrophy

Fibronectin (FN) is an extracellular matrix protein that binds to integrin receptors and couples cardiac myocytes to the basal lamina. Cardiac FN expression is elevated in models of pressure overload, and FN causes cultured cardiac myocytes to hypertrophy by a mechanism that has not been characterized in detail. In this study, we analyzed the gene expression changes induced by FN in purified rat neonatal ventricular myocytes using the Affymetrix RAE230A microarray, to understand how FN affects gene expression in cardiac myocytes and to separate the effects contributed by cardiac nonmyocytes in vivo. Pathway analysis using z-score statistics and comparison with a mouse model of cardiac hypertrophy revealed several pathways stimulated by FN in cardiac myocytes. In addition to the known cardiac myocyte hypertrophy markers, FN significantly induced metabolic pathways including virtually all of the enzymes of cholesterol biosynthesis, fatty acid biosynthesis, and the mitochondrial electron transport chain. FN also increased the expression of genes coding for ribosomal proteins, translation factors, and the ubiquitin-proteasome pathway. Interestingly, cardiac myocytes plated on FN showed elevated expression of the fibrosis-promoting peptides connective tissue growth factor (CTGF), WNT1 inducible signaling pathway protein 2 (WISP2), and secreted acidic cysteine-rich glycoprotein (SPARC). Our data complement in vivo studies and reveal several novel genes and pathways stimulated by FN, pointing to cardiac myocyte-specific mechanisms that lead to development of the hypertrophic phenotype.

Cell-cell contact between adult rat cardiac myocytes regulates Kv1.5 and Kv4.2 K+channel mRNA expression

1998 ◽  
Vol 275 (6) ◽  
pp. C1473-C1480 ◽  
Author(s):  
Kenneth M. Hershman ◽  
Edwin S. Levitan
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Cardiac Myocytes ◽  
Cell Density ◽  
Cardiac Myocyte ◽  
Live Cells ◽  
K Channel ◽  
Cell Contact ◽  
Adult Cardiac Myocytes ◽  
Cell Cell

Regulation of voltage-gated K+channel genes represents an important mechanism for modulating cardiac excitability. Here we demonstrate that expression of two K+channel mRNAs is reciprocally controlled by cell-cell interactions between adult cardiac myocytes. It is shown that culturing acutely dissociated rat ventricular myocytes for 3 h results in a dramatic downregulation of Kv1.5 mRNA and a modest upregulation of Kv4.2 mRNA. These effects are specific, because similar changes are not detected with other channel mRNAs. Increasing myocyte density promotes maintenance of Kv1.5 gene expression, whereas Kv4.2 mRNA expression was found to be inversely proportional to cell density. Conditioned culture medium did not mimic the effects of high cell density. However, paraformaldehyde-fixed myocytes were comparable to live cells in their ability to influence K+channel message levels. Thus the reciprocal effects of cell density on the expression of Kv1.5 and Kv4.2 genes are mediated by direct contact between adult cardiac myocytes. These findings reveal for the first time that cardiac myocyte gene expression is influenced by signaling induced by cell-cell contact.

scholarly journals Effects of tamoxifen inducible MerCreMer on gene expression in cardiac myocytes in mice

2022 ◽  
Author(s):  
Leila Rouhi ◽  
Siyang Fan ◽  
Sirisha M. Cheedipudi ◽  
Melis Olcum ◽  
Hyun-Hwan Jeong ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cardiac Function ◽  
Rna Sequencing ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Differentially Expressed ◽  
Therapeutic Interventions ◽  
Interferon Response ◽  
Sequencing Data ◽  
Dna Breaks

The Cre-LoxP technology, including the tamoxifen (TAM) inducible MerCreMer (MCM), is increasingly used to delineate gene function, understand the disease mechanisms, and test therapeutic interventions. We set to determine the effects of TAM-MCM on cardiac myocyte transcriptome. Expression of the MCM was induced specifically in cardiac myocytes upon injection of TAM to myosin heavy chain 6-MCM (Myh6-Mcm) mice for 5 consecutive days. Cardiac function, myocardial histology, and gene expression (RNA-sequencing) were analyzed 2 weeks after TAM injection. A total of 346 protein coding genes (168 up- and 178 down-regulated) were differentially expressed. Transcript levels of 85 genes, analyzed by a reverse transcription-polymerase chain reaction in independent samples, correlated with changes in the RNA-sequencing data. The differentially expressed genes were modestly enriched for genes involved in the interferon response and the tumor protein 53 (TP53) pathways. The changes in gene expression were relatively small and mostly transient and had no discernible effects on cardiac function, myocardial fibrosis, and apoptosis or induction of double-stranded DNA breaks. Thus, TAM-inducible activation of MCM alters cardiac myocytes gene expression, provoking modest and transient interferon and DNA damage responses without exerting other discernible phenotypic effects. Thus, the effects of TAM-MCM on gene expression should be considered in discerning the bona fide changes that result from the targeting of the gene of interest.

scholarly journals Cardiac myocyte exosomes: stability, HSP60, and proteomics

2013 ◽  
Vol 304 (7) ◽  
pp. H954-H965 ◽  
Author(s):  
Z. A. Malik ◽  
K. S. Kott ◽  
A. J. Poe ◽  
T. Kuo ◽  
L. Chen ◽  
...  
Keyword(s):  
Protein Content ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Lipid Vesicles ◽  
Underlying Mechanism ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Viral Particles ◽  
Adult Cardiac Myocytes ◽  
Cardiac Myocyte Apoptosis

Exosomes, which are 50- to 100-nm-diameter lipid vesicles, have been implicated in intercellular communication, including transmitting malignancy, and as a way for viral particles to evade detection while spreading to new cells. Previously, we demonstrated that adult cardiac myocytes release heat shock protein (HSP)60 in exosomes. Extracellular HSP60, when not in exosomes, causes cardiac myocyte apoptosis via the activation of Toll-like receptor 4. Thus, release of HSP60 from exosomes would be damaging to the surrounding cardiac myocytes. We hypothesized that 1) pathological changes in the environment, such as fever, change in pH, or ethanol consumption, would increase exosome permeability; 2) different exosome inducers would result in different exosomal protein content; 3) ethanol at “physiological” concentrations would cause exosome release; and 4) ROS production is an underlying mechanism of increased exosome production. We found the following: first, exosomes retained their protein cargo under different physiological/pathological conditions, based on Western blot analyses. Second, mass spectrometry demonstrated that the protein content of cardiac exosomes differed significantly from other types of exosomes in the literature and contained cytosolic, sarcomeric, and mitochondrial proteins. Third, ethanol did not affect exosome stability but greatly increased the production of exosomes by cardiac myocytes. Fourth, ethanol- and hypoxia/reoxygenation-derived exosomes had different protein content. Finally, ROS inhibition reduced exosome production but did not completely inhibit it. In conclusion, exosomal protein content is influenced by the cell source and stimulus for exosome formation. ROS stimulate exosome production. The functions of exosomes remain to be fully elucidated.

Microarray analysis of global changes in gene expression during cardiac myocyte differentiation

2002 ◽  
Vol 9 (3) ◽  
pp. 145-155 ◽  
Author(s):  
Chang-Fu Peng ◽  
Yi Wei ◽  
Jeffrey M. Levsky ◽  
Thomas V. McDonald ◽  
Geoffrey Childs ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cardiac Myocytes ◽  
Large Scale ◽  
Time Course ◽  
Cardiac Myocyte ◽  
Expression Patterns ◽  
Global Gene Expression ◽  
Global Changes ◽  
Novel Genes ◽  
Myocyte Differentiation

Significant progress has been made in defining pathways that mediate the formation of the mammalian heart. Little is known, however, about the genetic program that directs the differentiation of cardiac myocytes from their precursor cells. A major hindrance to this kind of investigation has been the absence of an appropriate cell culture model of cardiac myocyte differentiation. Recently, a subline of P19 cells (P19CL6) was derived that, following dimethyl sulfoxide (DMSO) treatment, differentiate efficiently over 10 days into spontaneously beating cardiac myocytes. We demonstrate that these cells are indeed cardiac myocytes as they express cell type-specific markers and exhibit electrophysiological properties indicative of cardiac myocytes. The requirement for DMSO stimulation in this paradigm was shown to be limited to the first 4 days, suggesting that critical events in the differentiation process occur over this interval. To uncover relationships among known genes and identify novel genes that mediate cardiac myocyte differentiation, a detailed time course of changes in global gene expression was carried out using cDNA microarrays. In addition to the activation of genes encoding cardiac transcription factors and structural proteins, increases were noted in the expression of multiple known genes and expressed sequence tags (ESTs). Analysis of the former suggested the involvement of a variety of signaling pathways in cardiac myocyte differentiation. The 16 ESTs whose expression was increased during the early, stimulus-dependent phase of cardiac myocyte differentiation may be novel regulators of this process. Thus this first report of large-scale changes in gene expression during cardiac myocyte differentiation has delineated relationships among the expression patterns of known genes and identified a number of novel genes that merit further study.

scholarly journals Signaling Pathways in Cardiac Myocyte Apoptosis

2016 ◽  
Vol 2016 ◽  
pp. 1-22 ◽  
Author(s):  
Peng Xia ◽  
Yuening Liu ◽  
Zhaokang Cheng
Keyword(s):  
Cardiovascular Diseases ◽  
Signaling Pathways ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Cardiomyocyte Apoptosis ◽  
Pharmacological Intervention ◽  
Future Research ◽  
Cardiac Myocyte Apoptosis

Cardiovascular diseases, the number 1 cause of death worldwide, are frequently associated with apoptotic death of cardiac myocytes. Since cardiomyocyte apoptosis is a highly regulated process, pharmacological intervention of apoptosis pathways may represent a promising therapeutic strategy for a number of cardiovascular diseases and disorders including myocardial infarction, ischemia/reperfusion injury, chemotherapy cardiotoxicity, and end-stage heart failure. Despite rapid growth of our knowledge in apoptosis signaling pathways, a clinically applicable treatment targeting this cellular process is currently unavailable. To help identify potential innovative directions for future research, it is necessary to have a full understanding of the apoptotic pathways currently known to be functional in cardiac myocytes. Here, we summarize recent progress in the regulation of cardiomyocyte apoptosis by multiple signaling molecules and pathways, with a focus on the involvement of these pathways in the pathogenesis of heart disease. In addition, we provide an update regarding bench to bedside translation of this knowledge and discuss unanswered questions that need further investigation.

Abstract 110: The Estrogen-related Receptor Gamma Promotes Maturation of iPSC-derived Cardiac Myocytes

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Tomoya Sakamoto ◽  
Teresa C Leone ◽  
Huei-Sheng Vincent Chen ◽  
Rick B Vega ◽  
Daniel P Kelly
Keyword(s):  
Gene Expression ◽  
Cardiac Myocytes ◽  
Cardiac Myocyte ◽  
Contractile Function ◽  
Neonatal Rat ◽  
Type I ◽  
Mature Adult ◽  
Gene Markers ◽  
Cardiac Contractile ◽  
Estrogen Related Receptor

The mammalian heart has a tremendous energy requirement to support normal contractile function. Recently, our group discovered that the estrogen-related receptor γ (ERRγ) coordinately controls oxidative energy metabolism and the type I fiber program in skeletal muscle. We hypothesized that this circuit is also operative in heart, serving to match capacity to produce ATP with energy utilizing processes such as contraction and ion transport. To identify downstream targets and pathways of ERRγ in cardiac myocytes, RNA-sequencing (RNA-seq) was conducted in neonatal rat ventricular myocytes (NRVMs) overexpressing ERRγ. ERRγ induced the expression of genes involved in mitochondrial oxidative phosphorylation, fatty acid oxidation, and the TCA cycle. Interestingly, ERRγ also induced expression of a broad program of adult cardiac contractile and structural genes including Myh6 (αMHC) and Ca 2+ handling genes such as Atp2a2 (SERCA2A). ERRγ-mediated activation of adult genes occurred concomitant with the suppression of fetal cardiac and embryonic/perinatal fast skeletal isoform contractile protein gene expression including Myh7 (βMHC), Myl1 and Myh3/8 . These results prompted us to explore the role of ERRγ in differentiation and maturation of human iPS-derived cardiac myocytes (iPSC-CM). The expression level of ERRγ was markedly upregulated during the transition to cardiac lineage commitment in parallel with induction of cardiogenic transcription factors (e.g. NKX2.5 and GATA4). Forced expression of ERRγ in iPSC-CM activated the broad adult cardiac myocyte program including oxidative metabolism and adult contractile gene expression. Of note, ERRγ increased MYH6 expression while decreasing expression of MYH7 and the fetal gene markers NPPA (ANF) and NPPB (BNP). The expression of TNNI3 , thought to be a marker of mature adult cardiomyocytes, was also induced by ERRγ. Conversely, CRISPR-Cas9n deletion of ERRγ in iPSC-CM resulted in diminished mitochondrial energy metabolic and adult cardiac contractile gene expression. These data provide evidence that ERRγ is a key nodal regulator of cardiac myocyte determination and terminal maturation. ERR signaling is a candidate target for driving iPSC-CM to the “adult” cardiac myocyte phenotype.

scholarly journals Focal Adhesion Kinase and p130Cas Mediate Both Sarcomeric Organization and Activation of Genes Associated with Cardiac Myocyte Hypertrophy

2001 ◽  
Vol 12 (8) ◽  
pp. 2290-2307 ◽  
Author(s):  
Branka Kovac̆ic̆-Milivojević ◽  
Frederick Roediger ◽  
Eduardo A.C. Almeida ◽  
Caroline H. Damsky ◽  
David G. Gardner ◽  
...  
Keyword(s):  
Gene Expression ◽  
Focal Adhesion Kinase ◽  
Cardiac Myocytes ◽  
Natriuretic Peptide ◽  
Focal Adhesion ◽  
Promoter Activity ◽  
Cardiac Myocyte ◽  
Gene Promoter ◽  
Myocyte Hypertrophy ◽  
Cardiac Myocyte Hypertrophy

Hypertrophic terminally differentiated cardiac myocytes show increased sarcomeric organization and altered gene expression. Previously, we established a role for the nonreceptor tyrosine kinase Src in signaling cardiac myocyte hypertrophy. Here we report evidence that p130Cas (Cas) and focal adhesion kinase (FAK) regulate this process. In neonatal cardiac myocytes, tyrosine phosphorylation of Cas and FAK increased upon endothelin (ET) stimulation. FAK, Cas, and paxillin were localized in sarcomeric Z-lines, suggesting that the Z-line is an important signaling locus in these cells. Cas, alone or in cooperation with Src, modulated basal and ET-stimulated atrial natriuretic peptide (ANP) gene promoter activity, a marker of cardiac hypertrophy. Expression of the C-terminal focal adhesion-targeting domain of FAK interfered with localization of endogenous FAK to Z-lines. Expression of the Cas-binding proline-rich region 1 of FAK hindered association of Cas with FAK and impaired the structural stability of sarcomeres. Collectively, these results suggest that interaction of Cas with FAK, together with their localization to Z-lines, is critical to assembly of sarcomeric units in cardiac myocytes in culture. Moreover, expression of the focal adhesion-targeting and/or the Cas-binding proline-rich regions of FAK inhibited ANP promoter activity and suppressed ET-induced ANP and brain natriuretic peptide gene expression. In summary, assembly of signaling complexes that include the focal adhesion proteins Cas, FAK, and paxillin at Z-lines in the cardiac myocyte may regulate, either directly or indirectly, both cytoskeletal organization and gene expression associated with cardiac myocyte hypertrophy.

Dynamic Changes in the Regulation of Energy Balance after Roux-en-Y Gastric Bypass Are Reflected in Time-Dependent Alterations in Liver Gene Expression

Diabetes ◽  
2018 ◽  
Vol 67 (Supplement 1) ◽  
pp. 289-LB
Author(s):  
M. AGOSTINA SANTORO ◽  
JOSEPH BRANCALE ◽  
JILL CARMODY GARRISON ◽  
SRIRAM MACHINENI ◽  
SCOTT A. LAJOIE ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gastric Bypass ◽  
Energy Balance ◽  
Time Dependent ◽  
Dynamic Changes ◽  
Liver Gene

Inhibition of the water channel aquaporin-1 prevents myocardial remodeling by blocking the transmembrane transport of hydrogen peroxide

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
V Montiel ◽  
R Bella ◽  
L Michel ◽  
E Robinson ◽  
J.C Jonas ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Cardiac Hypertrophy ◽  
Cardiac Myocytes ◽  
Cardiac Remodeling ◽  
Cardiac Myocyte ◽  
Water Channel ◽  
Transmembrane Transport ◽  
Funding Source ◽  
Water Pore

Abstract Background Pathological remodeling of the myocardium has long been known to involve oxidant signaling, but so far, strategies using systemic anti-oxidants have generally failed to prevent it. Aquaporins are a family of transmembrane water channels with thirteen isoforms currently known. Some isoforms have been implicated in oxidant signaling. AQP1 is the most abundant aquaporin in cardiovascular tissues but its specific role in cardiac remodeling remains unknown. Purpose We tested the role of AQP1 as a key regulator of oxidant-mediated cardiac remodeling amenable to targeted pharmacological therapy. Methods We used mice with genetic deletion of Aqp1 (and wild-type littermate), as well as primary isolates from the same mice and human iPSC/Engineered Heart Tissue to test the role of AQP1 in pro-hypertrophic signaling. Human cardiac myocyte-specific (PCM1+) expression of AQP's and genes involved in hypertrophic remodeling was studied by RNAseq and bioinformatic GO pathway analysis. Results RNA sequencing from human cardiac myocytes revealed that the archetypal AQP1 is a major isoform. AQP1 expression correlates with the severity of hypertrophic remodeling in patients with aortic stenosis. The AQP1 channel was detected at the plasma membrane of human and mouse cardiac myocytes from hypertrophic hearts, where it colocalizes with the NADPH oxidase-2 (NOX2) and caveolin-3. We show that hydrogen peroxide (H2O2), produced extracellularly, is necessary for the hypertrophic response of isolated cardiac myocytes and that AQP1 facilitates the transmembrane transport of H2O2 through its water pore, resulting in activation of oxidant-sensitive kinases in cardiac myocytes. Structural analysis of the amino acid residues lining the water pore of AQP1 supports its permeation by H2O2. Deletion of Aqp1 or selective blockade of AQP1 intra-subunit pore (with Bacopaside II) inhibits H2O2 transport in mouse and human cells and rescues the myocyte hypertrophy in human induced pluripotent stem cell-derived engineered heart muscle. This protective effect is due to loss of transmembrane transport of H2O2, but not water, through the intra-subunit pore of AQP1. Treatment of mice with clinically-approved Bacopaside extract (CDRI08) inhibitor of AQP1 attenuates cardiac hypertrophy and fibrosis. Conclusion We provide the first demonstration that AQP1 functions as an aqua-peroxiporin in primary rodent and human cardiac parenchymal cells. We show that cardiac hypertrophy is mediated by the transmembrane transport of H2O2 through the AQP1 water channel. Our studies open the way to complement the therapeutic armamentarium with specific blockers of AQP1 for the prevention of adverse remodeling in many cardiovascular diseases leading to heart failure. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): FRS-FNRS, Welbio

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